LOCUS       Exported                6306 bp ds-DNA     circular SYN 22-OCT-2018
DEFINITION  Corrected sequence.
            Added BglII (AGATCT) at 520 (upstream of synapsin promoter).
            
            Added BamHI (GGATCC) at 927 (downstream of synapsin promoter).
ACCESSION   .
VERSION     .
KEYWORDS    pAAV-EnhancedSynapsin-DIO-TVA950-EYFP-WPRE
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6306)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Nov 13, 2018 from SnapGene Viewer 4.2.6
            http://www.snapgene.com
COMMENT     Fumitaka 07.17.2013
            GT-AA-145
            For Deposit to Addgene
FEATURES             Location/Qualifiers
     source          1..6306
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     misc_feature    1..141
                     /label=L-ITR
     repeat_region   1..141
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     misc_feature    156..520
                     /label=CMV enhancer
     primer_bind     complement(179..200)
                     /label=Rev01
     enhancer        216..519
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     361..382
                     /label=For02
     misc_feature    528..927
                     /label=Synapsin Promoter
     primer_bind     complement(551..571)
                     /label=Rev02
     primer_bind     complement(588..607)
                     /label=Syn-Rev
     primer_bind     780..800
                     /label=For03
     misc_feature    952..985
                     /label=lox2722
     protein_bind    952..985
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GGATACTT). lox2272 sites are compatible with each
                     other, but incompatible with loxP or loxN sites."
     primer_bind     complement(986..1009)
                     /label=Rev03
     misc_feature    1036..1069
                     /label=loxP
     protein_bind    1036..1069
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     CDS             complement(1078..1797)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     exon            complement(1078..1794)
                     /label=EYFP (no start)
     primer_bind     complement(1089..1108)
                     /label=GFP (C' terminal CFP,YFP, or BFP)
     primer_bind     1179..1200
                     /label=For04
     primer_bind     complement(1386..1405)
                     /label=Rev04
     primer_bind     1582..1601
                     /label=For05
     primer_bind     1647..1666
                     /label=GFP Reverse
     primer_bind     complement(1786..1804)
                     /label=Rev05
     regulatory      1794..1803
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     misc_feature    complement(1810..2277)
                     /label=TVA codon opt
                     /label=TVA codon opt.
     primer_bind     1983..2003
                     /label=For06
     primer_bind     2164..2183
                     /label=TVA-For
     primer_bind     complement(2190..2209)
                     /label=Rev06
     primer_bind     complement(2233..2251)
                     /label=TVA-Rev
     regulatory      2277..2286
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     misc_feature    2278..2280
                     /label=TVA950-TAA
     misc_feature    complement(2291..2324)
                     /label=lox2722(1)
     protein_bind    complement(2291..2324)
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GGATACTT). lox2272 sites are compatible with each
                     other, but incompatible with loxP or loxN sites."
     misc_feature    complement(2375..2408)
                     /label=loxP(1)
     protein_bind    complement(2375..2408)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     primer_bind     2380..2409
                     /label=For07
     misc_feature    2423..3031
                     /label=WPRE
     misc_feature    2433..3021
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2486..2506)
                     /label=WPRE-R
     primer_bind     complement(2585..2609)
                     /label=Rev07
     primer_bind     2785..2804
                     /label=For08
     CDS             complement(2904..2915)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(2989..3008)
                     /label=Rev08
     polyA_signal    3051..3529
                     /label=hGH polyA
     polyA_signal    3053..3529
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     3189..3208
                     /label=For09
     primer_bind     complement(3252..3271)
                     /label=hGH-PA-R
     primer_bind     complement(3365..3385)
                     /label=Rev09
     primer_bind     3488..3508
                     /label=For10
     misc_feature    3569..3709
                     /label=R-ITR
     repeat_region   3569..3709
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     primer_bind     complement(3777..3797)
                     /label=Rev10
     rep_origin      3784..4239
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     rep_origin      3801..4107
                     /label=f1 Origin
     primer_bind     4375..4397
                     /label=pGEX Reverse
     promoter        4521..4625
                     /gene="bla"
                     /label=AmpR promoter
     CDS             4626..5486
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     misc_feature    4626..5483
                     /label=AmpR
     rep_origin      5634..6301
                     /label=pUC ori
     rep_origin      5657..6245
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6231..6251
                     /label=For01
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcaaag cccgggcgtc
       61 gggcgacctt tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag ggagtggcca
      121 actccatcac taggggttcc tgcggccgca cgcgttagtt attaatagta atcaattacg
      181 gggtcattag ttcatagccc atatatggag ttccgcgtta cataacttac ggtaaatggc
      241 ccgcctggct gaccgcccaa cgacccccgc ccattgacgt caataatgac gtatgttccc
      301 atagtaacgc caatagggac tttccattga cgtcaatggg tggagtattt acggtaaact
      361 gcccacttgg cagtacatca agtgtatcat atgccaagta cgccccctat tgacgtcaat
      421 gacggtaaat ggcccgcctg gcattatgcc cagtacatga ccttatggga ctttcctact
      481 tggcagtaca tctacgtatt agtcatcgct attaccatgg agatctctgc agagggccct
      541 gcgtatgagt gcaagtgggt tttaggacca ggatgaggcg gggtgggggt gcctacctga
      601 cgaccgaccc cgacccactg gacaagcacc caacccccat tccccaaatt gcgcatcccc
      661 tatcagagag ggggagggga aacaggatgc ggcgaggcgc gtgcgcactg ccagcttcag
      721 caccgcggac agtgccttcg cccccgcctg gcggcgcgcg ccaccgccgc ctcagcactg
      781 aaggcgcgct gacgtcactc gccggtcccc cgcaaactcc ccttcccggc caccttggtc
      841 gcgtccgcgc cgccgccggc ccagccggac cgcaccacgc gaggcgcgag ataggggggc
      901 acgggcgcga ccatctgcgc tgcggcggga tcctctagag tcgactccgg aataacttcg
      961 tataggatac tttatacgaa gttatgcaga atggtagctg gattgtagct gctattagca
     1021 atatgaaacc tcttaataac ttcgtatagc atacattata cgaagttatg gcgcgcctta
     1081 cttgtacagc tcgtccatgc cgagagtgat cccggcggcg gtcacgaact ccagcaggac
     1141 catgtgatcg cgcttctcgt tggggtcttt gctcagggcg gactggtagc tcaggtagtg
     1201 gttgtcgggc agcagcacgg ggccgtcgcc gatgggggtg ttctgctggt agtggtcggc
     1261 gagctgcacg ctgccgtcct cgatgttgtg gcggatcttg aagttcacct tgatgccgtt
     1321 cttctgcttg tcggccatga tatagacgtt gtggctgttg tagttgtact ccagcttgtg
     1381 ccccaggatg ttgccgtcct ccttgaagtc gatgcccttc agctcgatgc ggttcaccag
     1441 ggtgtcgccc tcgaacttca cctcggcgcg ggtcttgtag ttgccgtcgt ccttgaagaa
     1501 gatggtgcgc tcctggacgt agccttcggg catggcggac ttgaagaagt cgtgctgctt
     1561 catgtggtcg gggtagcggg cgaagcactg caggccgtag ccgaaggtgg tcacgagggt
     1621 gggccagggc acgggcagct tgccggtggt gcagatgaac ttcagggtca gcttgccgta
     1681 ggtggcatcg ccctcgccct cgccggacac gctgaacttg tggccgttta cgtcgccgtc
     1741 cagctcgacc aggatgggca ccaccccggt gaacagctcc tcgcccttgc tcaccatggt
     1801 ggcggccgcg gagaacaagt ctgcctggct cttgtcaggc accagcagct ccttggatgc
     1861 gctttcaaga ctgaagatgt cagacctgct ttttgctttg gacttccccc atgcagcgat
     1921 accacccaca gctaccaggc agcacaggag cactgcagtg atcagcatcc acatgcggcc
     1981 gtgattcctg gctggcagag cacgtccagg agcaggggca gtgggagcct ctgtgccgtt
     2041 gtccgtgggc accgcgggga tcgcgctggt cccgcagccc cactcgtccc gcccgtcgtc
     2101 gcagtcgggg tgtccgtcgc acagccagtc ctgcgggtaa cactccccgt gggcaccggg
     2161 cggctccgag cagcggaact gaccgggggg gcaacgggac aaagaaccgt taccggaccc
     2221 gttaccggtc acgttaccgg gcagcagcag cagcagcagc gcgggcagca gccgcgccat
     2281 ggtggctagc ataacttcgt ataaagtatc ctatacgaag ttatttgcct taacccagaa
     2341 attatcactg ttattcttta gaatggtgca aagaataact tcgtataatg tatgctatac
     2401 gaagttatga attcgatatc aagcttatcg ataatcaacc tctggattac aaaatttgtg
     2461 aaagattgac tggtattctt aactatgttg ctccttttac gctatgtgga tacgctgctt
     2521 taatgccttt gtatcatgct attgcttccc gtatggcttt cattttctcc tccttgtata
     2581 aatcctggtt gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg
     2641 tgtgcactgt gtttgctgac gcaaccccca ctggttgggg cattgccacc acctgtcagc
     2701 tcctttccgg gactttcgct ttccccctcc ctattgccac ggcggaactc atcgccgcct
     2761 gccttgcccg ctgctggaca ggggctcggc tgttgggcac tgacaattcc gtggtgttgt
     2821 cggggaaatc atcgtccttt ccttggctgc tcgcctatgt tgccacctgg attctgcgcg
     2881 ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc ggaccttcct tcccgcggcc
     2941 tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg agtcggatct
     3001 ccctttgggc cgcctccccg catcgatacc gagcgctgct cgagagatct acgggtggca
     3061 tccctgtgac ccctccccag tgcctctcct ggccctggaa gttgccactc cagtgcccac
     3121 cagccttgtc ctaataaaat taagttgcat cattttgtct gactaggtgt ccttctataa
     3181 tattatgggg tggagggggg tggtatggag caaggggcaa gttgggaaga caacctgtag
     3241 ggcctgcggg gtctattggg aaccaagctg gagtgcagtg gcacaatctt ggctcactgc
     3301 aatctccgcc tcctgggttc aagcgattct cctgcctcag cctcccgagt tgttgggatt
     3361 ccaggcatgc atgaccaggc tcagctaatt tttgtttttt tggtagagac ggggtttcac
     3421 catattggcc aggctggtct ccaactccta atctcaggtg atctacccac cttggcctcc
     3481 caaattgctg ggattacagg cgtgaaccac tgctcccttc cctgtccttc tgattttgta
     3541 ggtaaccacg tgcggaccga gcggccgcag gaacccctag tgatggagtt ggccactccc
     3601 tctctgcgcg ctcgctcgct cactgaggcc gggcgaccaa aggtcgcccg acgcccgggc
     3661 tttgcccggg cggcctcagt gagcgagcga gcgcgcagct gcctgcaggg gcgcctgatg
     3721 cggtattttc tccttacgca tctgtgcggt atttcacacc gcatacgtca aagcaaccat
     3781 agtacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga
     3841 ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg
     3901 ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat
     3961 ttagtgcttt acggcacctc gaccccaaaa aacttgattt gggtgatggt tcacgtagtg
     4021 ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata
     4081 gtggactctt gttccaaact ggaacaacac tcaaccctat ctcgggctat tcttttgatt
     4141 tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat
     4201 ttaacgcgaa ttttaacaaa atattaacgt ttacaatttt atggtgcact ctcagtacaa
     4261 tctgctctga tgccgcatag ttaagccagc cccgacaccc gccaacaccc gctgacgcgc
     4321 cctgacgggc ttgtctgctc ccggcatccg cttacagaca agctgtgacc gtctccggga
     4381 gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg cgcgagacga aagggcctcg
     4441 tgatacgcct atttttatag gttaatgtca tgataataat ggtttcttag acgtcaggtg
     4501 gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa
     4561 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga
     4621 agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc
     4681 ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg
     4741 gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc
     4801 gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat
     4861 tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg
     4921 acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag
     4981 aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa
     5041 cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc
     5101 gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca
     5161 cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc
     5221 tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc
     5281 tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg
     5341 ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta
     5401 tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag
     5461 gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga
     5521 ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc
     5581 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
     5641 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
     5701 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
     5761 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgtccttcta gtgtagccgt
     5821 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     5881 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     5941 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     6001 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
     6061 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
     6121 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
     6181 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
     6241 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
     6301 acatgt
//
